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Ligand-independent G protein-coupled estrogen receptor/G protein-coupled receptor 30 activity : Lack of receptor-dependent effects of G-1 and 17b-estradiol

Tutzauer, Julia LU orcid ; de Valdivia, Ernesto Gonzalez LU orcid ; Swärd, Karl LU ; Eilard, Ioannis Alexandrakis ; Broselid, Stefan LU ; Kahn, Robin LU ; Olde, Björn LU and Leeb-Lundberg, L. M.Fredrik LU (2021) In Molecular Pharmacology 100(3). p.271-282
Abstract

G protein-coupled receptor 30 (GPR30) is a membrane receptor reported to bind 17b-estradiol (E2) and mediate rapid nongenomic estrogen responses, hence also named G protein-coupled estrogen receptor. G-1 is a proposed GPR30-specific agonist that has been used to implicate the receptor in several pathophysiological events. However, controversy surrounds the role of GPR30 in G-1 and E2 responses. We investigated GPR30 activity in the absence and presence of G-1 and E2 in several eukaryotic systems ex vivo and in vitro in the absence and presence of the receptor. Ex vivo activity was addressed using the caudal artery from wild-type (WT) and GPR30 knockout (KO) mice, and in vitro activity was addressed using a HeLa cell line stably... (More)

G protein-coupled receptor 30 (GPR30) is a membrane receptor reported to bind 17b-estradiol (E2) and mediate rapid nongenomic estrogen responses, hence also named G protein-coupled estrogen receptor. G-1 is a proposed GPR30-specific agonist that has been used to implicate the receptor in several pathophysiological events. However, controversy surrounds the role of GPR30 in G-1 and E2 responses. We investigated GPR30 activity in the absence and presence of G-1 and E2 in several eukaryotic systems ex vivo and in vitro in the absence and presence of the receptor. Ex vivo activity was addressed using the caudal artery from wild-type (WT) and GPR30 knockout (KO) mice, and in vitro activity was addressed using a HeLa cell line stably expressing a synthetic multifunctional promoter (nuclear factor jB, signal transducer and activator of transcription, activator protein 1)-luciferase construct (HFF11 cells) and a human GPR30-inducible T-REx system (T-REx HFF11 cells), HFF11 and human embryonic kidney 293 cells transiently expressing WT GPR30 and GPR30 lacking the C-terminal PDZ (postsynaptic density-95/discs-large/zonula occludens-1 homology) motif SSAV, and yeast Saccharomyces cerevisiae transformed to express GPR30. WT and KO arteries exhibited similar contractile responses to 60 mM KCl and 0.3 mM cirazoline, and G-1 relaxed both arteries with the same potency and efficacy. Furthermore, expression of GPR30 did not introduce any responses to 1 mM G-1 and 0.1 mM E2 in vitro. On the other hand, receptor expression caused considerable ligand-independent activity in vitro, which was receptor PDZ motif-dependent in mammalian cells. We conclude from these results that GPR30 exhibits ligand-independent activity in vitro but no G-1- or E2-stimulated activity in any of the systems used. SIGNIFICANCE STATEMENT Much controversy surrounds 17b-estradiol (E2) and G-1 as G protein-coupled receptor 30 (GPR30) agonists. We used several recombinant eukaryotic systems ex vivo and in vitro with and without GPR30 expression to address the role of this receptor in responses to these proposed agonists. Our results show that GPR30 exhibits considerable ligand-independent activity in vitro but no G-1- or E2-stimulated activity in any of the systems used. Thus, classifying GPR30 as an estrogen receptor and G-1 as a specific GPR30 agonist is unfounded.

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organization
publishing date
type
Contribution to journal
publication status
published
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in
Molecular Pharmacology
volume
100
issue
3
pages
12 pages
publisher
American Society for Pharmacology and Experimental Therapeutics
external identifiers
  • scopus:85115780547
  • pmid:34330822
ISSN
0026-895X
DOI
10.1124/MOLPHARM.121.000259
language
English
LU publication?
yes
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Publisher Copyright: Copyright © 2021 by The Author(s) This is an open access article distributed under the CC BY-NC Attribution 4.0 International license.
id
754be55a-c139-41e0-8170-2e4bae7d569e
date added to LUP
2021-10-14 13:52:32
date last changed
2024-04-20 13:10:18
@article{754be55a-c139-41e0-8170-2e4bae7d569e,
  abstract     = {{<p>G protein-coupled receptor 30 (GPR30) is a membrane receptor reported to bind 17b-estradiol (E2) and mediate rapid nongenomic estrogen responses, hence also named G protein-coupled estrogen receptor. G-1 is a proposed GPR30-specific agonist that has been used to implicate the receptor in several pathophysiological events. However, controversy surrounds the role of GPR30 in G-1 and E2 responses. We investigated GPR30 activity in the absence and presence of G-1 and E2 in several eukaryotic systems ex vivo and in vitro in the absence and presence of the receptor. Ex vivo activity was addressed using the caudal artery from wild-type (WT) and GPR30 knockout (KO) mice, and in vitro activity was addressed using a HeLa cell line stably expressing a synthetic multifunctional promoter (nuclear factor jB, signal transducer and activator of transcription, activator protein 1)-luciferase construct (HFF11 cells) and a human GPR30-inducible T-REx system (T-REx HFF11 cells), HFF11 and human embryonic kidney 293 cells transiently expressing WT GPR30 and GPR30 lacking the C-terminal PDZ (postsynaptic density-95/discs-large/zonula occludens-1 homology) motif SSAV, and yeast Saccharomyces cerevisiae transformed to express GPR30. WT and KO arteries exhibited similar contractile responses to 60 mM KCl and 0.3 mM cirazoline, and G-1 relaxed both arteries with the same potency and efficacy. Furthermore, expression of GPR30 did not introduce any responses to 1 mM G-1 and 0.1 mM E2 in vitro. On the other hand, receptor expression caused considerable ligand-independent activity in vitro, which was receptor PDZ motif-dependent in mammalian cells. We conclude from these results that GPR30 exhibits ligand-independent activity in vitro but no G-1- or E2-stimulated activity in any of the systems used. SIGNIFICANCE STATEMENT Much controversy surrounds 17b-estradiol (E2) and G-1 as G protein-coupled receptor 30 (GPR30) agonists. We used several recombinant eukaryotic systems ex vivo and in vitro with and without GPR30 expression to address the role of this receptor in responses to these proposed agonists. Our results show that GPR30 exhibits considerable ligand-independent activity in vitro but no G-1- or E2-stimulated activity in any of the systems used. Thus, classifying GPR30 as an estrogen receptor and G-1 as a specific GPR30 agonist is unfounded.</p>}},
  author       = {{Tutzauer, Julia and de Valdivia, Ernesto Gonzalez and Swärd, Karl and Eilard, Ioannis Alexandrakis and Broselid, Stefan and Kahn, Robin and Olde, Björn and Leeb-Lundberg, L. M.Fredrik}},
  issn         = {{0026-895X}},
  language     = {{eng}},
  month        = {{09}},
  number       = {{3}},
  pages        = {{271--282}},
  publisher    = {{American Society for Pharmacology and Experimental Therapeutics}},
  series       = {{Molecular Pharmacology}},
  title        = {{Ligand-independent G protein-coupled estrogen receptor/G protein-coupled receptor 30 activity : Lack of receptor-dependent effects of G-1 and 17b-estradiol}},
  url          = {{http://dx.doi.org/10.1124/MOLPHARM.121.000259}},
  doi          = {{10.1124/MOLPHARM.121.000259}},
  volume       = {{100}},
  year         = {{2021}},
}